A comparison of non-viral vectors for gene delivery to pancreatic β-cells: Delivering a hypoxia-inducible vascular endothelial growth factor gene to rat islets

Byung Wan Lee, Hee Young Chae, Tran Thi Ngoc Tuyen, Dongchul Kang, Hyun Ah Kim, Minhyung Lee, Sung Hee Ihm

Research output: Contribution to journalArticle

11 Citations (Scopus)

Abstract

Although non-viral vectors are relatively safe, they have very low gene transfection efficiency, especially in pancreatic islet cells. To provide information on the use of nonviral vectors for transfecting genes into pancreatic islet cells, a comparative evaluation of non-viral options was performed. In vitro experiments were used to compare the transfection efficiency of three classes of non-viral vectors: Effectene, polyethylenimine (PEI, 25 kDa) and hemagglutinating virus of Japan-envelope (HVJ-E), into insulinoma cells (INS-1) and rat islets. Vascular endothelial growth factor (VEGF) gene with hypoxia-inducible RTP801 promoter was delivered into rat islets with Effectene and VEGF secretion under hypoxia was measured in the culture media. Luciferase activity and GFP assays indicated that Effectene exhibited the highest transfection efficiency, and HVJ-E was not suitable for transfection into pancreatic β-cells. The cytotoxicity of Effectene was found to be similar to that of 25-kDa PEI by 7-amino actinomycin D (7-AAD) flow cytometry and acridine orange/propidium iodide (AO/PI) assays. When RTP801 promoter-VEGF plasmid was delivered to rat islets with Effectene, VEGF secretion increased specifically in islets under hypoxia. In conclusion, Effectene showed higher gene-delivery efficiency for pancreatic islets compared with other classes of non-viral delivery systems and is promising as a gene delivery agent for pretransplant ex vivo gene therapy of islets.

Original languageEnglish
Pages (from-to)757-762
Number of pages6
JournalInternational journal of molecular medicine
Volume23
Issue number6
DOIs
Publication statusPublished - 2009 Jul 6

Fingerprint

Vascular Endothelial Growth Factor A
Islets of Langerhans
Transfection
Genes
Sendai virus
Polyethyleneimine
Acridine Orange
Insulinoma
Propidium
Luciferases
Genetic Therapy
Culture Media
Effectene
Hypoxia
Flow Cytometry
Plasmids

All Science Journal Classification (ASJC) codes

  • Genetics

Cite this

Lee, Byung Wan ; Chae, Hee Young ; Tuyen, Tran Thi Ngoc ; Kang, Dongchul ; Kim, Hyun Ah ; Lee, Minhyung ; Ihm, Sung Hee. / A comparison of non-viral vectors for gene delivery to pancreatic β-cells : Delivering a hypoxia-inducible vascular endothelial growth factor gene to rat islets. In: International journal of molecular medicine. 2009 ; Vol. 23, No. 6. pp. 757-762.
@article{ac80b43ef74248b88c9ed95f674e5479,
title = "A comparison of non-viral vectors for gene delivery to pancreatic β-cells: Delivering a hypoxia-inducible vascular endothelial growth factor gene to rat islets",
abstract = "Although non-viral vectors are relatively safe, they have very low gene transfection efficiency, especially in pancreatic islet cells. To provide information on the use of nonviral vectors for transfecting genes into pancreatic islet cells, a comparative evaluation of non-viral options was performed. In vitro experiments were used to compare the transfection efficiency of three classes of non-viral vectors: Effectene, polyethylenimine (PEI, 25 kDa) and hemagglutinating virus of Japan-envelope (HVJ-E), into insulinoma cells (INS-1) and rat islets. Vascular endothelial growth factor (VEGF) gene with hypoxia-inducible RTP801 promoter was delivered into rat islets with Effectene and VEGF secretion under hypoxia was measured in the culture media. Luciferase activity and GFP assays indicated that Effectene exhibited the highest transfection efficiency, and HVJ-E was not suitable for transfection into pancreatic β-cells. The cytotoxicity of Effectene was found to be similar to that of 25-kDa PEI by 7-amino actinomycin D (7-AAD) flow cytometry and acridine orange/propidium iodide (AO/PI) assays. When RTP801 promoter-VEGF plasmid was delivered to rat islets with Effectene, VEGF secretion increased specifically in islets under hypoxia. In conclusion, Effectene showed higher gene-delivery efficiency for pancreatic islets compared with other classes of non-viral delivery systems and is promising as a gene delivery agent for pretransplant ex vivo gene therapy of islets.",
author = "Lee, {Byung Wan} and Chae, {Hee Young} and Tuyen, {Tran Thi Ngoc} and Dongchul Kang and Kim, {Hyun Ah} and Minhyung Lee and Ihm, {Sung Hee}",
year = "2009",
month = "7",
day = "6",
doi = "10.3892/ijmm_00000189",
language = "English",
volume = "23",
pages = "757--762",
journal = "International Journal of Molecular Medicine",
issn = "1107-3756",
publisher = "Spandidos Publications",
number = "6",

}

A comparison of non-viral vectors for gene delivery to pancreatic β-cells : Delivering a hypoxia-inducible vascular endothelial growth factor gene to rat islets. / Lee, Byung Wan; Chae, Hee Young; Tuyen, Tran Thi Ngoc; Kang, Dongchul; Kim, Hyun Ah; Lee, Minhyung; Ihm, Sung Hee.

In: International journal of molecular medicine, Vol. 23, No. 6, 06.07.2009, p. 757-762.

Research output: Contribution to journalArticle

TY - JOUR

T1 - A comparison of non-viral vectors for gene delivery to pancreatic β-cells

T2 - Delivering a hypoxia-inducible vascular endothelial growth factor gene to rat islets

AU - Lee, Byung Wan

AU - Chae, Hee Young

AU - Tuyen, Tran Thi Ngoc

AU - Kang, Dongchul

AU - Kim, Hyun Ah

AU - Lee, Minhyung

AU - Ihm, Sung Hee

PY - 2009/7/6

Y1 - 2009/7/6

N2 - Although non-viral vectors are relatively safe, they have very low gene transfection efficiency, especially in pancreatic islet cells. To provide information on the use of nonviral vectors for transfecting genes into pancreatic islet cells, a comparative evaluation of non-viral options was performed. In vitro experiments were used to compare the transfection efficiency of three classes of non-viral vectors: Effectene, polyethylenimine (PEI, 25 kDa) and hemagglutinating virus of Japan-envelope (HVJ-E), into insulinoma cells (INS-1) and rat islets. Vascular endothelial growth factor (VEGF) gene with hypoxia-inducible RTP801 promoter was delivered into rat islets with Effectene and VEGF secretion under hypoxia was measured in the culture media. Luciferase activity and GFP assays indicated that Effectene exhibited the highest transfection efficiency, and HVJ-E was not suitable for transfection into pancreatic β-cells. The cytotoxicity of Effectene was found to be similar to that of 25-kDa PEI by 7-amino actinomycin D (7-AAD) flow cytometry and acridine orange/propidium iodide (AO/PI) assays. When RTP801 promoter-VEGF plasmid was delivered to rat islets with Effectene, VEGF secretion increased specifically in islets under hypoxia. In conclusion, Effectene showed higher gene-delivery efficiency for pancreatic islets compared with other classes of non-viral delivery systems and is promising as a gene delivery agent for pretransplant ex vivo gene therapy of islets.

AB - Although non-viral vectors are relatively safe, they have very low gene transfection efficiency, especially in pancreatic islet cells. To provide information on the use of nonviral vectors for transfecting genes into pancreatic islet cells, a comparative evaluation of non-viral options was performed. In vitro experiments were used to compare the transfection efficiency of three classes of non-viral vectors: Effectene, polyethylenimine (PEI, 25 kDa) and hemagglutinating virus of Japan-envelope (HVJ-E), into insulinoma cells (INS-1) and rat islets. Vascular endothelial growth factor (VEGF) gene with hypoxia-inducible RTP801 promoter was delivered into rat islets with Effectene and VEGF secretion under hypoxia was measured in the culture media. Luciferase activity and GFP assays indicated that Effectene exhibited the highest transfection efficiency, and HVJ-E was not suitable for transfection into pancreatic β-cells. The cytotoxicity of Effectene was found to be similar to that of 25-kDa PEI by 7-amino actinomycin D (7-AAD) flow cytometry and acridine orange/propidium iodide (AO/PI) assays. When RTP801 promoter-VEGF plasmid was delivered to rat islets with Effectene, VEGF secretion increased specifically in islets under hypoxia. In conclusion, Effectene showed higher gene-delivery efficiency for pancreatic islets compared with other classes of non-viral delivery systems and is promising as a gene delivery agent for pretransplant ex vivo gene therapy of islets.

UR - http://www.scopus.com/inward/record.url?scp=67649515538&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=67649515538&partnerID=8YFLogxK

U2 - 10.3892/ijmm_00000189

DO - 10.3892/ijmm_00000189

M3 - Article

C2 - 19424601

AN - SCOPUS:67649515538

VL - 23

SP - 757

EP - 762

JO - International Journal of Molecular Medicine

JF - International Journal of Molecular Medicine

SN - 1107-3756

IS - 6

ER -